Single-Cell Virology: On-Chip Investigation of Viral Infection Dynamics
Feng Guo,
Sixing Li,
Mehmet Umut Caglar,
Zhangming Mao,
Wu Liu,
Andrew Woodman,
Jamie J. Arnold,
Claus O. Wilke,
Tony Jun Huang,
Craig E. Cameron
Affiliations
Feng Guo
Department of Engineering Science and Mechanics, The Pennsylvania State University, University Park, PA 16802, USA
Sixing Li
Molecular Cellular and Integrative Biosciences Graduate Program, The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA
Mehmet Umut Caglar
Center for Computational Biology and Bioinformatics, Institute for Cellular and Molecular Biology, and Department of Integrative Biology, The University of Texas at Austin, Austin, TX 78712, USA
Zhangming Mao
Department of Engineering Science and Mechanics, The Pennsylvania State University, University Park, PA 16802, USA
Wu Liu
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA
Andrew Woodman
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA
Jamie J. Arnold
Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA
Claus O. Wilke
Center for Computational Biology and Bioinformatics, Institute for Cellular and Molecular Biology, and Department of Integrative Biology, The University of Texas at Austin, Austin, TX 78712, USA
Tony Jun Huang
Department of Engineering Science and Mechanics, The Pennsylvania State University, University Park, PA 16802, USA; Molecular Cellular and Integrative Biosciences Graduate Program, The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA; Corresponding author
Craig E. Cameron
Molecular Cellular and Integrative Biosciences Graduate Program, The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA; Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA; Corresponding author
Summary: We have developed a high-throughput, microfluidics-based platform to perform kinetic analysis of viral infections in individual cells. We have analyzed thousands of individual poliovirus infections while varying experimental parameters, including multiplicity of infection, cell cycle, viral genotype, and presence of a drug. We make several unexpected observations masked by population-based experiments: (1) viral and cellular factors contribute uniquely and independently to viral infection kinetics; (2) cellular factors cause wide variation in replication start times; and (3) infections frequently begin later and replication occurs faster than predicted by population measurements. We show that mutational load impairs interaction of the viral population with the host, delaying replication start times and explaining the attenuated phenotype of a mutator virus. We show that an antiviral drug can selectively extinguish the most-fit members of the viral population. Single-cell virology facilitates discovery and characterization of virulence determinants and elucidation of mechanisms of drug action eluded by population methods. : Guo et al. use a microfluidics device installed on a fluorescence microscope to monitor the kinetics of viral infection in single cells. Between-cell variation in outcomes of infection exists at all phases of the life cycle. Cellular gene expression governs the eclipse phase; viral genetics govern replication rate and yield. Keywords: viral infection, virus-host interaction, temporal dynamics, single-cell analysis, stochastic gene expression, microfluidics, live-cell imaging
